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Team:Mexico-UNAM-CINVESTAV/Notebook/Week Five
From 2010.igem.org
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'''We prepare all to repeat ligation; PCR2 with Psb1C3 and PCR4 with Psb1C3''' | '''We prepare all to repeat ligation; PCR2 with Psb1C3 and PCR4 with Psb1C3''' | ||
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| + | ==''Thursday''== | ||
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| + | '''Plan''' | ||
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| + | '''We did ligations (Fast ligation one hour) Pcr 2 with Psb1C3 and PCR 4 with Psb1C3 then transformed DH5α''' | ||
| + | |||
| + | '''competent cells, and incubated at 37º overnight.''' | ||
Revision as of 04:48, 26 October 2010
Week #5
04th October - 08 October 2010
Monday
- Let's picked up colonys our transformant cells and prepare all to do miniprep.
For this we used 30ml de LB liquid agar medium per falcon tube at 80μ/ml
Cloramphenicol concentration we left overnight culture to 37º
Tuesday
With a overnight inoculum we did a miniprep.
The miniprep was done with a Quiagen Kit.
Then we going to do a duble digest with EcoRI and PstI to check out our ligations.
| DNA | 5μl |
| Buffer NB2 | 1μl |
| EcoRI | 0.5μl |
| PstI | 0.5μl |
| H2O | 2.7μl |
| BSA | 0.3μl |
| Total | 10μl |
Wensday
We run a gel to confirm our ligations as follow
- PCR 1 Lig S with Psb1C3 Confirmed
- PCR 2 Lig S with Psb1C3 Unconfirmed
- PCR 3 Lig S with Psb1C3 Confirmed
- PCR 4 Lig C with Psb1C3 Unconfirmed
We prepare all to repeat ligation; PCR2 with Psb1C3 and PCR4 with Psb1C3
Thursday
Plan
We did ligations (Fast ligation one hour) Pcr 2 with Psb1C3 and PCR 4 with Psb1C3 then transformed DH5α
competent cells, and incubated at 37º overnight.
